Uso de peptidases para produção de hidrolisado hipoalergênico do leite bovino

Abstract

Cow’s milk allergy is a global health problem that generally affect infants and children. The disorder in most cases is transitory, however it must be managed to avoid development issues. Hydrolyzed formulas are the most suitable alternative to cow’s milk. Thus, the present study aimed to study the peptidases from laticifer plants and insect midgut able to perform cow’s milk proteins hydrolysis. The hydrolysis assays were performed varying the following parameters: enzyme:substrate ratio, pH and time of reaction. In addition, it was measured the degree of hydrolysis and the residual antigenicity of the hydrolysates. The serine peptidases purified from Danaus plexippus midgut (DpGp) were able to perform caseins hydrolysis at 1:25 (enzyme:substrate ratio), after 4 hours of reaction at pH 9.0. Using the same parameters, whey proteins hydrolysis were performed after 24 hours; however the hydrolysis of pre-heated whey proteins (85 °C for 30 min) caused a reduction in the time of reaction as well as in its antigenicity. The immunoenzymatic assays showed that milk hydrolyzed by DpGp exhibited residual antigenicity similar to commercial partially hydrolyzed formulae. The latex peptidases from Calotropis procera (CpLP), Cryptostegia grandiflora (CgLP) e Carica papaya (CapLP) were able to perform caseins hydrolysis at 1:225 (enzyme:substrate ratio), after 30 min of reaction at pH 6.5. Only CapLP hydrolyzed the whey proteins after 24 hours of reaction. The heat denaturation of the whey proteins also caused an enhance in the degree of hydrolysis and reduced the residual antigenicity of the hydrolysates. The cow’s milk hydrolyzed by CpLP trigged immune reactions in cow’s milk allergy mouse model. However, the milk hydrolyzed by CgLP and CapLP did not elicited clinical symptoms after oral administration in sensitized mice. The peptidases of DpGp, CgLP and CapLP are promising enzymes to develop novel hypoallergenic formulas for children with milk allergy